Construction Of Yeast Two Hybrid CDNA Library Of Maize (Zea Mays L.) Endosperm And BD-vector

Maize is an important food, feed crop and raw materials in industry in the world. We have made a great progress in maize traditional breeding. The advent of plant gene engineering and advanced molecular techniques for plant breeding provides powerful tools for genetically improving maize as to enhance its resistance to biotic stress and grain quality. The yeast two-hybrid system, will now make it much easier for interested researchers come to grips with the technology. The yeast two-hybrid system can be applied to the analysis of protein-protein, protein-RNA and protein-small organic ligand interactions.For this reason, the following text is devoted to cDNA library construction. The first chain of cDNA was synthesized by reverse transcript from maize endosperm mRNA as template by SMART methods. The ds cDNA was amplified by long distance-PCR in existence of DNA polymerase. The purified ds cDNA and the linearized plasmid pGADT7-Rec were co-transformed into the competent yeast AH109. Recombinants plasmids formed through homologous recombination in vivo. All the transformants were harvested from the SD/-Leu plates and then constituted the maize endosperm cDNA library. 1.8×10~8 transformants were obtained from the maize endosperm cDNA library. In conclusion, the yeast two-hybrid cDNA library of maize endosperm in yeast cell AH109 was successfully constructed.Starch is not only the main source of food but also is one of the important raw materials in industry. Therefore it has been one of the key researches in genetic breeding to improve starch quality. Therefore it has been one of the key researches in genetic breeding to improve starch quality. Since ss plays an extremely vital role in determining the structure formation of starch, it is helpful to understand biosynthesis mechanism of starch in order to thoroughly study it. So we use RT-PCR to clone the cDNA of maize gene ss1 from maize endosperm total RNA with the primers designed according to the sequence of gene ss1 in the GenBank. A DNA-BD fusion vector was constructed using the cDNA of ss1.